d-Xylulose Fermentation to Ethanol by Saccharomyces cerevisiae

We used commercial bakers' yeast (Saccharomyces cerevisiae) to study the conversion of d-xylulose to ethanol in the presence of d-xylose. The rate of ethanol production increased with an increase in yeast cell density. The optimal temperature for d-xylulose fermentation was 35 degrees C, and the optimal pH range was 4 to 6. The fermentation of d-xylulose by yeast resulted in the production of ethanol as the major product; small amounts of xylitol and glycerol were also produced. The production of xylitol was influenced by pH as well as temperature. High pH values and low temperatures enhanced xylitol production. The rate of d-xylulose fermentation decreased when the production of ethanol yielded concentrations of 4% or more. The slow conversion rate of d-xylulose to ethanol was increased by increasing the yeast cell density. The overall production of ethanol from d-xylulose by yeast cells under optimal conditions was 90% of the theoretical yield.     

Mechanism of Fermentation Conversion from Polyalcohol Fermentation to Ethanol Fermentation by Pichia miso

A possible mechanism of fermentation conversion is described from polyalcohol fermentation to ethanol fermentation by Pichia miso. Little alcohol dehydrogenase activity was found in polyalcohol-producing cells, whereas higher enzyme activity was induced by ethanol-producing cells. The fermentation conversion may be caused by the different levels of alcohol dehydrogenase activity between polyalcohol- and ethanol-producing cells. It was also shown that yeast growth was inhibited and that yeast cells were lysed by ethanol (at 6g/100ml) that accumulated in 24 hr.     

Specific Features of Fermentation of D-Xylose and D-Glucose by Xylose-Assimilating Yeasts

The ability to assimilate D-glucose and D-xylose was studied in 21 yeast species of the following genera: Candida, Kluyveromyces, Pachysolen, Pichia, and Torulopsis. All the cultures fermented D-glucose with the formation of ethanol. During the assimilation of D-xylose, ethanol was produced by P. stipitis and C. shehatae, whereas xylitol was produced by C. didensiae, C. intermediae, C. parapsilosis, C. silvanorum, C. tropicalis, K. fragilis, K. marxianus, P. guillermondii, and T. molishiama. The yeast P. tannophilus produced comparable amounts of both alcohols. The possible use of xylose-assimilating yeasts for the production of xy-litol and ethanol is discussed.     

生淀粉高浓度酒精发酵的研究

本研究利用国内常用的糖化酶制剂糖化生玉米面中的淀粉,同时接种酵母菌,在30℃下,探讨了玉米淀粉的高浓度酒精发酵工艺。选择到了一株产高浓度酒精酵母菌,H0菌株。发酵温度为30℃、pH4一s、加糖化酶量为每克原料300单位、酵母接种量3％(v／v)和原料加量为33．0％(w／v)时,这株酵母菌在70小时内可产生17．5％(v／v)的乙醇。如果原料加量为36．0％(w／v)时,该菌株在96小时内可以产生18．O％(v／v)的乙醇。在前一种加料情况下,成熟发酵醪中的pH为5、残还原糖为O．19％、残总糖为3．5“。在后一种加料情况下,成熟发酵醪中的pH为5、残还原糖为0．81％、残总糖为5．1％。     

休哈塔假丝酵母乙醇发酵性能的研究

木糖的高效发酵是制约纤维素燃料乙醇生产的技术瓶颈之一,高性能发酵菌种的开发是本领域研究的重点。以木糖发酵的典型菌株休哈塔假丝酵母为材料,研究氮源配比、葡萄糖和木糖初始浓度、葡萄糖添加及典型抑制物等因素对其木糖利用和乙醇发酵性能的影响规律。结果表明,硫酸铵更适宜于木糖和葡萄糖发酵产乙醇。在摇瓶振荡发酵条件下,该酵母可发酵164.0 g/L葡萄糖生成61.9 g/L乙醇,糖利用率和乙醇得率分别为99.8%和74.0%;受酵母细胞膜上转运体系的限制,对木糖的最高发酵浓度为120.0 g/L,可生成45.7 g/L乙醇,糖利用率和乙醇得率分别达到94.8%和87.0%。休哈塔假丝酵母发酵木糖的主要产物为乙醇,仅生成微量的木糖醇;添加葡萄糖可促进木糖的利用;休哈塔假丝酵母在葡萄糖发酵时的乙酸和甲酸的耐受浓度分别为8.32和2.55 g/L,木糖发酵时的乙酸和甲酸的耐受浓度分别为6.28和1.15 g/L。     

Direct Fermentation of d-Xylose to Ethanol by Kluyveromyces marxianus Strains

Eight strains of Kluyveromyces marxianus were screened, and all of them were found to ferment the aldopentose d-xylose directly to ethanol under aerobic conditions. One of these strains, K. marxianus SUB-80-S, was grown in a medium containing 20 g of d-xylose per liter, and the following results were obtained: maximum ethanol concentration, 5.6 g/liter; ethanol yield, 0.28 g of ethanol per g of d-xylose (55% of theoretical); maximum specific growth rate, 0.12 h; 100% d-xylose utilization was completed in 48 h.     

Fermentation of d-Xylose to Ethanol by Genetically Modified Klebsiella planticola

d-Xylose is a plentiful pentose sugar derived from agricultural or forest residues. Enteric bacteria such as Klebsiella spp. ferment d-xylose to form mixed acids and butanediol in addition to ethanol. Thus the ethanol yield is normally low. Zymomonas spp. and most yeasts are unable to ferment xylose, but they do ferment hexose sugars to ethanol in high yield because they contain pyruvate decarboxylase (EC 4.1.1.1), a key enzyme that is absent from enteric bacteria. This report describes the fermentation of d-xylose by Klebsiella planticola ATCC 33531 bearing multicopy plasmids containing the pdc gene inserted from Zymomonas mobilis. Expression of the gene markedly increased the yield of ethanol to 1.3 mol/mol of xylose, or 25.1 g/liter. Concurrently, there were significant decreases in the yields of formate, acetate, lactate, and butanediol. Transconjugant Klebsiella spp. grew almost as fast as the wild type and tolerated up to 4% ethanol. The plasmid was retained by the cells during at least one batch culture, even in the absence of selective pressure by antibiotics to maintain the plasmid. Ethanol production was 31.6 g/liter from 79.6 g of mixed substrate per liter chosen to simulate hydrolyzed hemicellulose. The physiology of the wild-type of K. planticola is described in more detail than in the original report of its isolation.     

Fermentation of d-Xylose and l-Arabinose to Ethanol by Erwinia chrysanthemi

Erwinia spp. are gram-negative facultative anaerobes within the family Enterobacteriacae which possess several desirable traits for the conversion of pentose sugars to ethanol, such as the ability to ferment a broad range of carbohydrates and the ease with which they can be genetically modified. Twenty-eight strains of Erwinia carotovora and E. chrysanthemi were screened for the ability to ferment d-xylose to ethanol. E. chrysanthemi B374 was chosen for further study on the basis of its superior (4%) ethanol tolerance. We have characterized the fermentation of d-xylose and l-arabinose by the wild type and mutants which bear plasmids containing the pyruvate decarboxylase gene from Zymomonas mobilis. Expression of the gene markedly increased the yields of ethanol (from 0.7 up to 1.45 mol/mol of xylose) and decreased the yields of formate, acetate, and lactate. However, the cells with pyruvate decarboxylase grew only one-fourth as fast as the wild type and tolerated only 2% ethanol. Alcohol tolerance was stimulated by the addition of yeast extract to the growth medium. Xylose catabolism was characterized by a high saturation constant K(s) (4.5 mM).     

休哈塔假丝酵母发酵木糖制乙醇过程研究

木质纤维素原料水解产物的主要成分是葡萄糖和木糖,其中葡萄糖很容易发酵,致使木糖成为木质纤维素发酵的关键,休哈塔假丝酵母(Candida shehatae)1766是自然界木糖发酵性能较好的天然酵母之一。研究了发酵温度、发酵时间、接种量、初始pH值、摇床转速等因素对休哈塔假丝酵母1766发酵木糖生产乙醇的影响,由正交试验初步确定了休哈塔假丝酵母发酵木糖制乙醇工艺的适宜条件为好氧条件,发酵时间为2d,发酵温度为28℃,摇床转速为150r/min,初始pH值为5,此时乙醇收率最高可达68.62%。     

Aerobic degradation of diethyl phthalate by Sphingomonas sp

An aerobic diethyl phthalate (DEP) degrading bacterium, DEP-AD1, was isolated from activated sludge. Based on its 16S rDNA sequence, this isolate was identified belonging to Sphingomonas genus with 99% similarity to Sphingomonas sp. strain C28242 and 98% similarity to S. capsulate. The specific degradation rate of DEP was concentration dependent with a maximum of 14 mg-DEP/(Lh). Results of degradation tests showed that DEP-AD1 could also degrade monoethyl phthalate (MEP), dimethyl phthalate (DMP), dibutyl phthalate (DBP), and diethylhexyl phthalate (DEHP), but not phthalate and benzoate.     

高强度酒精连续发酵

从多株酒精酵母菌株中筛选出S.cerevisiaeKG作为酒精生产菌株。该菌株具有较好的酒精发酵活力和絮凝性。使用具有细胞循环的连续发酵双罐系统,酒精生产强度达到30.5（g／1.h）。根据该系统的动力学模型,讨论了获得高强度酒精连续发酵的途径。     

Invasive infections due to Clavispora lusitaniae

Three cases of Clavispora lusitaniae invasive fungal infections are reported. All three infections appeared in cancer patients presented with fungaemia, one additionally with meningitis. Two of them were breakthrough -- they developed during therapy with conventional amphotericin B with a dose of 0.5 mg kg(-1) day(-1) . All three were cured: two with intravenous fluconazol and one with an increasing dose (1 mg kg(-1) day(-1)) of amphotericin B. In one of two breakthrough cases the sensitivity of the strain to antifungals was tested against antifungal agents and showed in vitro resistance to amphotericin B (MIC 2 eta g ml(-1)).     

Production of Ethanol from Maltose by Zymobacter palmae Fermentation

The production of ethanol from maltose by Zymobacter palmae T109 in monoculture fermentations, and in co-culture fermentations together with Zymomonas mobilis B69 was studies. Zymobacter palmae T109, produced 5.5% (w/v) of ethanol when co-cultured with Zymomonas mobilis B69, but Zymobacter palmae T109 produced only 4.9% (w/v) ethanol from 15% (w/v) maltose medium in monoculture fermentation.     

蔗渣水解液发酵乙醇的研究

研究了酵母（Ｐｉｃｈｉａｓｔｉｐｉｔｉｓ）Ｙ７１２４在限制供氧条件下尽管反应初期葡萄糖消耗速率大于木糖,但在一定时间后,葡萄糖的消耗速率变慢,而木糖消耗速率变快直至耗尽的现象。建立了气升柱以Ｐ．Ｓｔｉｐｉｔｉｓ转化木糖为目的和以溢流柱Ｓａｃｈａｒｏｍｙｃｅｓｃｅｒｉｖｉｓｉａｅ转化残留葡萄糖为目的的串联发酵乙醇工艺,即流加５倍浓缩的蔗渣水解液,Ｄ＝０.１ｈ^－１,还原糖总利用率为９７.２％,酒精浓度为４６.５ｇ／Ｌ,生产率为４.１ｇ／Ｌ·ｈ。     

Fundamental Studies on the Aerobic Fermentation

The oxygen uptake rate of aggregated mycelia is decreased to an extent which, in the case of a typical spherical aggregate, could be estimated depending on its diameter, mycelial density, oxygen diffusivity, and so forth. Equations were presented in this paper to evaluate the oxygen uptake rate of an mold pellet. A favorable agreement was found between the calculation and the experiment.     

代谢工程改造运动发酵单胞菌用于提高乙醇产量

目的:采用可以在运动发酵单胞菌中表达的操纵子构建重组运动发酵单胞菌,用于提高该细菌对高温高糖的耐受性和提高乙醇产量.方法:用外来的YfdZ、MetB和Hsp构建的多顺反子质粒,转化运动发酵单胞菌而使其获得新的代谢途径.在玉米水解液中,验证了该多顺反子质粒对运动发酵单胞菌产生乙醇的影响.结果:与对照菌相比,在37℃和糖浓度为28%的培养条件下,该基因工程菌的乙醇产量提高到183.2%.在37℃,糖浓度为28%并添加氮源的条件下,该基因工程菌的乙醇产量提高到148.0%.结论:YfdZ、MetB及Hsp三种基因的共同作用能显著提高运动发酵单胞菌的乙醇产量和发酵温度.     

代谢工程改善野生酵母利用木糖产乙醇的性能

从256个自然样品中筛选得到1株可高效转化D-木糖的酵母。通过生理生化和分子生物学方法鉴定, 证实该菌株是属于Candida tropicalis。以该酵母为研究对象, 增加木糖醇脱氢酶表达量, 通过改变代谢流以达到提高酒精产率的目的。以pXY212-XYL2质粒为基础载体, 构建了含有潮霉素抗性的pYX212-XYL2-Hygro, 电击转化进入野生型C. tropicalis, 潮霉素抗性筛选, 得到含高拷贝木糖醇脱氢酶基因的重组菌株C. tropicalis XYL2-7。重组菌的比酶活达到0.5 u/mg protein, 比原始菌株提高了3倍。实验表明, 重组菌木糖醇得率比原始菌株降低了3倍, 酒精得率提高了5倍。首次通过实验验证了热带假丝酵母利用木糖产乙醇的可行性, 这对研究酵母利用秸秆、麦糠、谷壳等纤维质农业废弃物生产燃料乙醇具有重要启示。     

代谢工程改善野生酵母利用木糖产乙醇的性能

从256个自然样品中筛选得到1株可高效转化D-木糖的酵母。通过生理生化和分子生物学方法鉴定, 证实该菌株是属于Candida tropicalis。以该酵母为研究对象, 增加木糖醇脱氢酶表达量, 通过改变代谢流以达到提高酒精产率的目的。以pXY212-XYL2质粒为基础载体, 构建了含有潮霉素抗性的pYX212-XYL2-Hygro, 电击转化进入野生型C. tropicalis, 潮霉素抗性筛选, 得到含高拷贝木糖醇脱氢酶基因的重组菌株C. tropicalis XYL2-7。重组菌的比酶活达到0.5 u/mg protein, 比原始菌株提高了3倍。实验表明, 重组菌木糖醇得率比原始菌株降低了3倍, 酒精得率提高了5倍。首次通过实验验证了热带假丝酵母利用木糖产乙醇的可行性, 这对研究酵母利用秸秆、麦糠、谷壳等纤维质农业废弃物生产燃料乙醇具有重要启示。     

Efficient and Direct Fermentation of Starch to Ethanol by Sake Yeast Strains Displaying Fungal Glucoamylases

Aspergillus oryzae glucoamylases encoded by glaA and glaB, and Rhizopus oryzae glucoamylase, were displayed on the cell surface of sake yeast Saccharomyces cerevisiae GRI-117-UK and laboratory yeast S. cerevisiae MT8-1. Among constructed transformants, GRI-117-UK/pUDGAA, displaying glaA glucoamylase, produced the most ethanol from liquefied starch, although MT8-1/pUDGAR, displaying R. oryzae glucoamylase, had the highest glucoamylase activity on its cell surface.